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Method for localization of sialic acid on cell surface and cell interior in peripheral blood: A pilot study
Author(s) -
Pönniö Maritta,
Soukka Jori,
Sillanaukee Pekka,
Franck Johan,
Seveus Lahja
Publication year - 2004
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20132
Subject(s) - intracellular , fluorescein isothiocyanate , glycocalyx , fluorescein , lectin , cell , confocal microscopy , sialic acid , fluorophore , lysis , fluorescence microscope , confocal , chemistry , cell membrane , cytoplasm , biophysics , biochemistry , microbiology and biotechnology , biology , fluorescence , physics , geometry , mathematics , quantum mechanics
This article presents a method for identification and localization of cell surface and intracellular sialoglycoconjugates of peripheral blood cells. To reveal cell surface conjugates, a sample of peripheral blood was incubated with lectin after centrifugation and rinsing. For intracellular localization in leukocytes, RBCs were lysed and the membranes were permeabilized prior to cytochemical reaction. Fluorescein isothiocyanate conjugated lectins were used for visualization in fluorescence microscope. All lectins bound specifically to the surface of erythrocytes. Confocal microscopy showed surface and intracellular labeling of permeabilized leukocytes. A part of the signal in eosinophils originated from binding of anionic fluorophore to cationic granular proteins. Microsc. Res. Tech. 65:292–294, 2004. © 2005 Wiley‐Liss, Inc.